title: Fragment screening by weak affinity chromatography: comparison with established techniques for screening against HSP90.
abstract: The increasing use of fragment-based lead discovery (FBLD) in industry as well as in academia creates a high demand for sensitive and reliable methods to detect the binding of fragments to act as starting points in drug discovery programs. Nuclear magnetic resonance (NMR), surface plasmon resonance (SPR), and X-ray crystallography are well-established methods for fragment finding, and thermal shift and fluorescence polarization (FP) assays are used to a lesser extent. Weak affinity chromatography (WAC) was recently introduced as a new technology for fragment screening. The study presented here compares screening of 111 fragments against the ATPase domain of HSP90 by all of these methods, with isothermal titration calorimetry (ITC) used to confirm the most potent hits. The study demonstrates that WAC is comparable to the established methods of ligand-based NMR and SPR as a hit-id method, with hit correlations of 88% and 83%, respectively. The stability of HSP90 WAC columns was also evaluated and found to give 90% reproducibility even after 207 days of storage. A good correlation was obtained between the various technologies, validating WAC as an effective technology for fragment screening.
authors.affiliation: Department of Chemistry and Biomedical Sciences, Linnaeus University, Kalmar, Sweden.
authors.forename: Elinor
authors.initials: E
authors.lastname: Meiby
authors.forename: Heather
authors.initials: H
authors.lastname: Simmonite
authors.forename: Loic
authors.initials: L
authors.lastname: le Strat
authors.forename: Ben
authors.initials: B
authors.lastname: Davis
authors.forename: Natalia
authors.initials: N
authors.lastname: Matassova
authors.forename: Jonathan D
authors.initials: JD
authors.lastname: Moore
authors.forename: Michael
authors.initials: M
authors.lastname: Mrosek
authors.forename: James
authors.initials: J
authors.lastname: Murray
authors.forename: Roderick E
authors.initials: RE
authors.lastname: Hubbard
authors.forename: Sten
authors.initials: S
authors.lastname: Ohlson
mesh_terms: D000595:Amino Acid Sequence; D002846:Chromatography, Affinity / Q000379:methods*; D018360:Crystallography, X-Ray; D004353:Drug Evaluation, Preclinical / Q000379:methods; D018841:HSP90 Heat-Shock Proteins / Q000032:analysis* / Q000235:genetics; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000032:analysis* / Q000235:genetics; D017434:Protein Structure, Tertiary
chemical_list: D018841:HSP90 Heat-Shock Proteins; D010446:Peptide Fragments